Abstract
We have developed an in vitro immunization protocol of human peripheral blood mononuclear cells (PBMC) for generating human antigen-specific antibodies. By using this protocol, B cells producing antigen-specific antibody can be propagated within a week. In the present study, we tried to establish an efficient strategy to clone variable region genes of antigen-specific human monoclonal antibody by applying in vitro immunized PBMC to the phage display method. To evaluate the efficiency of our strategy, variable region genes prepared by PCR from PBMC immunized in vitro with mite-extract (ME) and non-immunized PBMC were applied to the phage display method. After concentrating the ME-specific phage antibody by several rounds of panning, the number of phage antibodies specific for ME and antigen specificity both increased by using in vitro immunized PBMC. These results demonstrate that efficiency in acquisition of variable region genes of antigen specific human monoclonal antibody can be improved by in vitro immunizing PBMC used as a source of antibody genes.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
